Bactericidal, germicidal, and antiseptic materials



Patented July 21, 1942 UNITED STATES PATENT OFFICE BACTERICIDAL, GERMICIDAL AND ANTISEPTIC MATERIALS Albert K. Epstein and Benjamin E. Harris, Chicago, Ill.

No Drawing. Application October 4, 1940,

Serial No. 359,720]

18 Claims.

This invention relates to antiseptic, bactericidal, bacteriostatic and germicidal materials and compositions. I

It has been found that certain materials, hereinafter described in detail, possess antiseptic, bactericidal (including bacteriostatic) and germicidal properties to a marked degree and are substantially non-toxic toward even highly delicate animal tissue.

At least many of the antiseptic, bactericidal, bacteriostatic and germicidal materials of the present invention, which are employed particularly in the form of their hydrochlorides, or the like, as hereinafter set forth, fall within the scope of the general formula:

Where R is an organic radical containing at least six and preferably from 8 to 18 carbon atoms, X and Y are hydrogen, alkyl, cycloalkyl, alkoxyl, aralkyl, aryl, or alkylol, and n is zero or a small whole numbers The radical B- may be of aliphatic, cycloaliphatic, aromatic, or aromatic-aliphatic character, and may contain substituent groups such as amino, hydroxy, halogen, sulphate, sulphonic, phosphate, oarboxyl, nitrile, and the like, as will be pointed out hereinafter, but it is particularly preferred that it be unsubstituted aliphatic or fatty and contain at least eight carbon atoms.

The antiseptic, bactericidal, bacteriostatic and germicidal materials of the present invention 1) 20 grams. of alanine were dissolved in 60 cc. of hot water and the resulting solution was treated with 350 cc. of isopropanol. The mass was filtered and the precipitate was dried at 100 degrees C. for 4 /2 hours.

(2) 17 grams of the dried, purified alanine, produced in part 1 hereof, were suspended in 350 cc. of acetyl chloride at degrees C. To the resulting suspension, 58 grams of fresh PCls (dry and powdered) were added in two portions, and the mixture was stirred at 0 degrees C. for minutes and then the stirring was continued for an additional hour at room temperature. 2 grams of PC were then added and the stirring was continued for a further- 1% hours. The mass was then allowed to stand for several hours at room temperature. The mixture was then stirred for /2 hour and filtered, and the precipitate was washed with acetyl chloride and ligroin. Most of the ligroin was removed from the precipitate in vacuo over P205.

(3) The precipitate, produced in part 2 hereof, was added to 100 grams of lauryl alcohol and the resulting suspension was heated at 100 de grees C. {for /2 hour whereupon evolution of HCl occurred and a homogeneous solution was obtained. About 200 cc. of heroin were then added, the mass was cooled in an ice bath until crystallization occurred, the precipitate was filtered off and washed several times with a total of 1500 cc. of cold ligroin. It was then suspended in 100 cc. of benzene at about 25 degrees derived from various sources.

tate was a white, crystalline material, having the formula omwmm-cm-o-o-on-cn.

0 manner (4) The precipitate produced in part 3 hereof was freed of benzene in vacuo over P205, hydrochloric acid being liberated. After 1 hour in vacuo, there was an HCl odor and, after an additional A; hour over P205, the HCl odor had disappeared. The product was water-soluble and alcohol soluble.

Example B (1) 25 grams of alpha-amino iso-butyric acid were dissolved in 150 cc. of hot water and 1 liter of isopropanol was added. The precipitate was collected, dried at 100 degrees C. for 4 hours, powdered and sieved.

(2) 21 grams of the dried, powdered and sieved, purified alpha-amino iso-butyric acid, produced in part 1 hereof, were suspended in 400 cc. of acetyl chloride, and 60 grams of dry, powdered PO15 were added in two portions, over a period oi 15 minutes, at 0 degrees C. The resulting suspension was then stirred for 2 hours at room temperature, 18 grams of P015 were added, and the stirring was continued for /2 hour. The mixture was allowed to stand for several hours, it was then stirred for 1 hour, and filtered, and the resulting precipitate was washed with acetyl chloride and ligroin.

(3) The amino acid hydrochloride, produced in part 2 hereof, was mixed with 100 grams of lauryl alcohol and the resulting mixture was heated in an oil bath. At degrees C., a vigorous evolution of HCl occurred and a waterwhite solution was produced. The solution was then maintained at 70 degrees C.- degrees C. for 15 minutes. 1 liter of ligroin was then added; the precipitate which formed was collected and then suspended in and washed with 1 liter of ligroin. The white, crystalline precipitate was placed in a vacuum desiccator over P205, the vacuum being maintained for a time suflicient to remove the ligroin and HCl. 60 grams of a product were obtained which was completely soluble in water and which had the following formula The hydrochloride of the lauryl alcohol ester of glycine was prepared in the manner described in steps 2, 3 and 4 of Example A, using pure glycine in place of alanine.

The higher molecular weight organic radical represented by R in the general formula may be Among such sources may be mentioned the straight chain and branched chain higher molecular weight alcohols, saturated and unsaturated, of aliphatic, aromatic or aliphatic-aromatic character including,'among others, hexyl alcohol, heptyl alcohol, octyl alcohol, nonyl alcohol, decyl alcohol, undecyl alcohol, lauryl alcohol, myristyl alcohol, cetyl alcohol,

voleyl alcohol, linoleyl alcohol, stearyl alcohol,

dro n Wasv addedthereto. The precipi- 75 and the like; branched chain octyl, decyl, dodecyl,

cholesterol,

branched chain alcohols. Preferably, the alcohols which are utilized are those corresponding to the fatty acids occurring in triglyceride oils and fats of vegetable or animal origin, natural or hydrogenated, such as corn oil, cottonseed oil, sesame oil, coconut oil, palm kernel oil, sunflower seed oil, lard, tallow, soya bean oil and the like, those alcohols containing from 12 to 18 carbon atoms being preferred. Other alcohols which may be employed are the cyclo-aliphatic or alicyclic alcohols such as the sterols, as, for example,

iso-cholesterol, phytosterol, sito- -sterol, hydroaromatic alcohols such as abietol, and such unsaturated alcohols as linalool, citronellol, geraniol and the like and hydrogenated products-of the foregoing. Also included within the class of alcohols which maybe employed are a such compounds as the hydroxy and alpha-hydroxy higher aliphatic and fatty acids as, for example, ricinoleic acid, alpha-hydroxy stearic acid,. alpha-hydroxy lauric acid, di-hydroxy stearic acid, i-hydroxy-stearic acid, alpha-hydroxy palmitic acid, and the like, as well as esters of hydroxy-fatty acids, such as ethyl ricinoleate,

castor oil, butyl alpha-hydroxystearate, cetylhydroxystearate, and the like. Still other alcohol molecules are the carboxylic acid amides of alkylolamines such as lauric acid amide of monoethanolamine, oleic acid amide of mono-propanolamine, and the like.

The term alcohols, as employed herein, is intended to include alcohols which may or may not contain other groups such as carboxylic, carbonyl, cyanogen, sulphone, sulphoxide, halogen, amine, amide, sulphonic, sulphate, or other radicals.

The alcohols obtainable by substituting alkyl or acyl radicals, preferably of high molecular weight, in place of the hydrogen of one or more hydroxy groups of polyhydroxy substances or polyhydric alcohols, it being understood that at least one hydroxy group attached to the nucleus of the polyhydroxy substance or polyhydric alcohol remains, are also within the scope of the alcohols from which the materials ofthe present invention may be produced. As examples of such alcohols may be mentioned partially esterified or partially etherified mono-, di-, and poly-saccharides, and sugar alcohols, such as monolauric acid ester of sucrose, monostearie acid ester of dextrose, monopalmitic acid ester of sorbitol,"

of diethylene glycol, monolauric acid esterof ethylene glycol, monolauric acid ester of triethanolamine, monostearic acid ester of tripropanolamine, and the like. The aliphatic polyhydroxy substance which is partially esterified or etherified to provide one of the reacting constituents herein may be selected from a large class andincludes, among those mentioned, glycerol; glycols such as ethylene glycol, propylene glycol, trimethylene glycol, butylene glycol and the like;.polyglycols such as diethylene glycol;

pentaerythritol; quercitol; dihydroxy acetone; triethanolamine; tripropanolamine; polyglycerols such as diglycerol, triglycerol, tetraglycerol and the like including mixtures thereof; carbohydrates and sugars including mono, diand polysaccharides, such as dextrose, sucrose, xylose, arabinose, galactose, fructose, maltose, mannose, dextrin', starch, and the like; the natural and synthetic, simple and complex glucosides; sugar (For convenience, all hydroxyl groups are written facing one way). In general, this type of alcohol maybe represented by the formula wherein R is a radical selected from the group consisting of alkyl, cyclo-alkvl, aryl, aralkyl, and acyl, and substitution products thereof, X is the residue of the aliphatic polyhydroxy substance,

and v and w are small whole numbers.

It is, of course, obvious that the alcohols from which the materials disclosed herein may be produced may be prepared in accordance with any desired method. For example, many of these alcohols may be prepared by the so-called Bouveault and Blane method or, alternately, by the reduction or catalytic reaction with hydrogen of natural or hydrogenated animal or vegetable fats and oils, or mixtures thereof, in accordance with well known'practices. Again the .al-

cohols may be derived from synthetic processes such as by the oxidation of hydrocarbons or may be prepared by saponification of waxes and the like. Alternatively, they may be prepared by re-:

oil or the free fatty acids of coconut oil. Lauryl alcohol comprises about 45% of the total alcohol mixture, the remaining alcohols running from C6 to C18. Again, mixtures of alcohols such as are present in the so-c'alled sperm oil alcohols, as

. Sarcinu lutea well as those present in wool-fat, may efficaciously be utilized. Indeed, these higher molecular weight alcohols are generally offered on the market in the form of mixtures of different alcohols. If desired, special fractions which predominate in a certain particular higher molecular weight alcohol may be utilized or, if so desired, the maetrials may be prepared from a single, substantially pure alcohol.

The amino-carboxylic acids which are or may be utilized in the preparation of the materials include, among those previously mentioned and others, alpha amino acetic acid, alpha amino propionic acid, alpha amino butyric acid, beta amino butyric acid, alpha amino-n-valeric acid, alpha amino isobutyric acid, alpha amino isovaleric acid, phenylalanine, tyrosine, tryptophane, cystine, arginine, amino poly-carboxylic acids such as amino succinic acid, amino glutaric acid, amino citric acid, amino malonic acid, amino adipic acid, amino sebacic acid, and the like; the amino-carboxylic acids or mixtures thereof obtained by the hydrolysis of proteins or proteincontaining materials such as gluten, soya bean flour and the like, whether by enzymatic action, by acids or by any other means. Other aminocarboxlic acids include the synthetic polypeptides as well as those resulting from the hydrolysis of proteins or protein-containing materials. Illustrative examples of such polypeptides are glycylglycine and leucyl-glycyl-glycine. It will be understood that mixtures of any two or more of the foregoing or other amino-carboxylic acids may be utilized in the preparation of the antiseptic, bactericidal and germicidal agents of the present invention. ,It will also be understood that any of these amino-carboxylic acids may be esterifled with any of the'alcohols disclosed hereinabove.

The materials are used, as previously indicated, preferably in the form of their hydrochlorides. They may also be used, in certain instances, in the form of their sulphates, phosphates, nitrates and, in general, in the form of salts with preferably relatively strong inorganic acids.

As indicative of the unusual bactericidal and germicidal effects produced with the materials vals, the following concentrations of the compound set forth hereinabove were employed: 1/100; 1/500; 1/1000; 1/2500; 1/5000; 1/10,000; and 1/ 15,000. The phenol was employed in concentrations of 1/60; 1/70; 1/80; and l/90. In the following concentrations, growth was observed in 5 minutes but none in 10 minutes:

Compound set forth hereinabove 1/500 Phenol 1/ 65 The phenol coeiiicient of the compound under discussion was approximately 8.

In toxicity tests, by intraperitoneal injections using white mice weighing approximately 20 grams, the animals survived the injection of 200 to 300 mg.' of the compound under discussionper kilogram of body weight. Toxicity tests on the tissues of the rabbit eye showed that one to three drops of a 0.1% physiological saline solution of the compound under discussion, applied twice each day for a period of four days, produced no reaction. On prolonged irrigation, continuously for five minutes twice each day, in a 0.1% concentration, slight immediate reddening of the eye occurred, clearing up completely by the following morning. After four days of such treatment, there was no evidence of inflammation or damage in the eye. In further tests, the eye was irrigated continuously for five minutes once each day with a 0.5% solution of the compound under discussion. This treatment caused an immediate reddening and some edema of the conjunctival membranes, but the reaction cleared up completely in 24 hours. A continuation of the treatment for five days did not cause permanent reactions.

Tests conducted on mice infected with virulent cultures of pneumococcus, by injecting various dilutions of the virulent organisms intraperitoneally simultaneously with the compound under discussion, showed a definite measure of protection against as high as 100,000 times the lethal dose of pneumococcus.

The inhibitory action on bacterial metabolism varies with the number of carbon atoms, with the chain length, and with the configuration of the amino-carboxylic acid and alcohol portions of the present invention, reference may be had,

Concentrations of 0.03% gave 90% to 100% in hibition of bacterial metabolism with suspensions containing approximately five billion cells per cc.,

in the usual bufier solutions, tested in the Warburg apparatus, of the following micro-organisms:

Staphylococcus aureus' Staphylococcus albus (Gram positive) (Gram positive) (Gram positive) (Gram positive) Y (Gram negative) (Gram negative) Micrococcus tetragenus Bacilli coli Aerobacter aerogenes Ps. pyocyaneus (Gram negative) Proteus vulgan's (Gram negative) At the lower concentrations, of the order of 0.003%, a strong inhibitory eflect was observed on some but not on all of the bacteria.

In theF. D. A. method for determination of phenol coeflicients, employing Staphylococcus aureus at 3'7 degrees C., at Ste 10 minute inter of the molecule. For example, the compound of Example B is more effective than that of Example A and considerably more efi'ective than that of the hydrochloride of the lauryl alcohol ester of alpha-amino acetic acid.

As indicated hereinabove, the compounds are best used in the form of salts, preferably in the form of substantially neutral salts or at a pH of about 6.7. The hydrochlorides are especially satisfactory but other salts may be employed such as the hydrobromides, hydrlodides, lactates, tartrates, citrates, sulphates, phosphates, and the like. The free esters of the amino-carboxylic acids generally possess limited solubility in water and, particularly for that reason, the esters are used in the form of their salts, the salts possessing substantially greater water-solubility. It is, however, within the broader scope of the invention to employ the free esters, using solvent media other than water or water alone, or by solubilizing the esters in any desired manner.

It will be appreciated that the numerous compounds disclosed herein will vary somewhat in bactericidal, germicidal, and antiseptic power or potency and that the effects will also vary depending upon the particular micro-organisms or bacteria or the like or the environment in which the compounds are used. The compounds are employed particularly in dilute aqueous or modiwherein R is an organic radical containing at fied aqueous solutions, usually on the order of 0.1% to several thousandths of one percent. They may be used as such or in conjunction with other agents having bactericidal, germicidal, or antiseptic action.

Wherever the term higher is employed as referring to higher molecular weight organic acids or the like, it will be understood to cover compounds or radicals having at least six carbon 1 atoms unless otherwise specifically stated.

wherein R. is an organic radical containing at least six'carbon atoms, X and Y are selected from the group consisting of hydrogen, alkyl. cycloalkyl, alkoxyl, aralkyl, aryl, and alkylol, and n is zero or a small whole number.

2. Bactericidal, germicidal and antiseptic agents comprising substances having the formula wherein R is an organic radical containing at least six carbon atoms, X andY are selected from the group consisting of hydrogen, alkyl, cycloalkyl, alkoxyl, aralkyl, aryl, and alkylol, and n is zero or a small whole number.

3. Bactericidal, germicidal and antiseptic agents comprising salts of substances having the formula n-o-o-orkwmnr-cm I NH:

wherein R is an organic containing from eight to eighteen carbon atoms, Y is selected from the group consisting of hydrogen, alkyl, cycloalkyl, alkoxyl, aralkyl, aryl, and alkylol, and n is zero or a small whole number.

4. Bactericidal, germicidal and antiseptic agents comprising salts of esters of alcohols having at least six carbon atoms with amino-carboxylic acids.

5. Bactericidal, germicidal and antiseptic agents comprising water-soluble inorganic acid salts of estersof aliphatic alcohols having at least six carbon atoms with aliphatic amino carboxylic acids.

6. Bactericidal, germicidal agents comprising hydrochlorides of esters of alcohols having at least six carbon atoms with amino carboxylic acids.

'l. Bactericidal, germicidal and antiseptic compositions comprising a dilute aqueous solution of v a salt of an ester of an alcohol, having at least six carbon atoms, with an amino carboxylic acid.

8. Bactericidal, germicidal and antiseptic compositions comprising a dilute aqueous solution of a substantially neutral salt of a substance corresponding to the formula least six carbon atoms, X and Y are selected from the group consisting of hydrogen, alkyl, cycloalkyl, alkoxyl, aralkyl, aryl, and alkylol, and n is zero or a small whole number.

9. Bactericidal, germicidal and antiseptic compositions comprising a dilute aqueous solution of a substance corresponding to the formula R-0-c-cucnY),.c1n

Nmlrol wherein R is an aliphatic radical containing at least six carbon atoms, Y is selected from the and antiseptic group consisting of hydrogen, alkyl, cycloalkyl, alkoxyl,,aralkyl, aryl, and alkylol, ora small whole number.

l0. Bactericidal, germicidal and antiseptic compositions comprisinga dilute aqueous solution of a substance corresponding to the formula wherein R is an organic radical containing at least six carbon atoms, X and Y are selected from the group consisting of hydrogen, alkyl, cycloalkyl, alkoxyl, aralkyl, aryl, and alkylol, and n is zero or a small whole number. v

,11. Bactericidal. germicidal and antiseptic compositions comprising a dilute aqueous solution of a substantially neutral salt of an ester of aliphatic alcohols containing from eight to eighteen carbon atoms with alpha-amino aliphatic carboxylic acids.

14. Bactericidal, germicidal and antiseptic agents comprising hydrochlorides ofesters of lauryl alcohol with alpha-amino acetic acid.

15. Bactericidal, germicidal and antiseptic agents comprising salts of esters of lauryl alcohol with alpha-amino aliphatic carboxylic acids containing not more than eight carbon atoms.

16. Bactericidal, germicidal and antiseptic agents comprising esters of alcohols having at least six carbon atoms with amino carboxylic;

acids.

17. Bactericidal,- germicidal and antiseptic ALBERT K. EPS'IEIN. BENJAMIN R. HARRIS.

and n is zero I 

